摘要
目的建立环介导等温扩增(loop-mediated isothermal amplification, LAMP)方法快速检测食品中的过敏原牡蛎成分。方法根据国家生物技术信息中心的牡蛎线粒体序列,通过Primer Explorer version 5.0软件设计引物并筛选出LAMP特异性扩增引物。并进一步对反应体系优化,对该方法的灵敏度、特异性以及稳定性进行验证。对10种牡蛎阳性样品、14种阴性样品、4类牡蛎相关食品进行检测。结果该方法可以检测出含牡蛎成分0.1%, DNA浓度为0.01 ng/μL的样品。在实验时间上大幅缩短,反应可在25～45 min内结束,并且可以在微量体系下完成,对于食品相关产品的检出率为100%。结论该方法操作简单、成本较低、特异性高、灵敏度好,适用于食品中过敏原牡蛎成分的检测。
Objective To establish a method for rapid detection of food allergens oyster ingredients by loop-mediated isothermal amplification(LAMP).Methods According to the mitochondrial sequences of oysters from the national biotechnology information center,primers were designed by using Primer Explorer version 5.0 software,and LAMP specific amplification primers were selected.Then the reaction system was optimized,and the sensitivity,specificity and stability of the method were verified.Totally 10 kinds of oyster positive samples,14 kinds of negative samples and 4 kinds of oyster related food were detected.Results The method could detect the samples with 0.1%oyster composition and DNA concentration of 0.01 ng/μL.The test time was greatly shortened,could be completed within 25 to 45 min,and could be completed under a trace system,with the detection rate of 100%for food-related products.Conclusion The method is simple operation,low cost,high specificity and high sensitivity,and is suitable for detecting allergen oyster components in foods.
作者
张懿翔
于媛媛
宋春宏
李妍
熊薇
葛宇
ZHANG Yi-Xiang;YU Yuan-Yuan;SONG Chun-Hong;LI Yan;XIONG Wei;GE Yu(Shanghai Institute of Quality Inspection and Technical Research,Shanghai 200233,China)
出处
《食品安全质量检测学报》
CAS
2019年第7期1804-1810,共7页
Journal of Food Safety and Quality
基金
上海市技术性贸易措施应对专项项目(16TBT004)
上海市科学技术委员会科研计划项目(16DZ0501700)
上海市食品质量安全检测与评价专业技术服务平台(18DZ2292400)~~
